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951.
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Ratul Saha Robert S. Donofrio Susan T. Bagley 《Journal of industrial microbiology & biotechnology》2010,37(8):843-848
A TaqMan quantitative real-time polymerase chain reaction (qPCR) assay was developed for the detection and enumeration of
three Pseudomonas species belonging to the mendocina sublineage (P. oleovorans, P. pseudoalcaligenes, and P. oleovorans subsp. lubricantis) found in contaminated metalworking fluids (MWFs). These microbes are the primary colonizers and serve as indicator organisms
of biodegradation of used MWFs. Molecular techniques such as qPCR are preferred for the detection of these microbes since
they grow poorly on typical growth media such as R2A agar and Pseudomonas isolation agar (PIA). Traditional culturing techniques not only underestimate the actual distribution of these bacteria but
are also time-consuming. The primer–probe pair developed from gyrase B (gyrB) sequences of the targeted bacteria was highly sensitive and specific for the three species. qPCR was performed with both
whole cell and genomic DNA to confirm the specificity and sensitivity of the assay. The sensitivity of the assay was 101 colony forming units (CFU)/ml for whole cell and 13.7 fg with genomic DNA. The primer–probe pair was successful in determining
concentrations from used MWF samples, indicating levels between 2.9 × 103 and 3.9 × 106 CFU/ml. In contrast, the total count of Pseudomonas sp. recovered on PIA was in the range of <1.0 × 101 to 1.4 × 105 CFU/ml for the same samples. Based on these results from the qPCR assay, the designed TaqMan primer–probe pair can be efficiently
used for rapid (within 2 h) determination of the distribution of these species of Pseudomonas in contaminated MWFs. 相似文献
954.
955.
Most of the previous studies on the effects of iron deficiency on skeletal muscle respiratory capacity and work performance have been investigated in severe or moderate iron-deficiency anemia. We report here that even in mild iron deficiency where the hemoglobin concentration was 10 g/dl and the iron stores in livers and spleen were not completely depleted, a marked reduction in succinate dehydrogenase was observed in skeletal muscles but not in heart. Similarly, cytochrome oxidase activities were reduced. Although no significant change in glycerophosphate dehydrogenase was detected in the iron-deficient rats, exposure to cold in this group greatly reduced this enzyme activity. As cold acclimatization accelerates marrow erythropoiesis (20) which in turn, demands more iron, it seems that in the iron-insufficient state, this iron demand for marrow activity may persist at the expense of the tissue iron pool, resulting in a marked reduction in glycerophosphate dehydrogenase activities. Since succinate dehydrogenase plays a significant role in the impairment of mitochondrial function and early fatigue of iron-deficient muscle (11), the present study shows that even in mild iron deficiency, some loss of muscle functions could result as succinate dehydrogenase activities were greatly reduced. 相似文献
956.
957.
By freezing blocks of paraffin-embedded tissues to a convenient temperature it is possible to obtain routinely 1 micron sections that can be further processed as normal thicker sections. Normal and disposable steel knives can be used and the staining time should be increased in most procedures. Gradual freezing of blocks to the temperature of dry ice is the simplest and safest way to obtain an adequate temperature. The best results were obtained using as fixative 4% paraformaldehyde in phosphate buffered saline solution. 相似文献
958.
Summary The sex circle model for crossing over and gene conversion proposed by Stahl has a number of properties which are in conflict with a considerable body of recombination data in fungi. The model is unable to explain the observed frequencies of gene conversion and postmeiotic segregation for particular mutants in several species. It does not provide an explanation for fine structure map expansion, nor does it account satisfactorily for the polarised distribution of outside markers amongst allelic recombinants. 相似文献
959.
960.
The chemical synthesis of a tetradecadeoxyribonucleotide, d-EtSp(A-T-G-G-A-A-A-C-T-G-C-G-G-C), is described. This oligomer, designated Fragment 4δ, constitutes the 5′-terminus of the plus strand of a projected duplex coding for S-Peptide2–14 derived from Ribonuclease A. The Fragment was constructed by block condensation via a phosphorothioate anchor. Complications due to inadvertent phosphotriester condensations are discussed. Arguments justifying the sequence selection are presented. 相似文献